Department of Joint Surgery, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
Department of Orthopedics, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, China.
Int J Rheum Dis. 2020 Aug;23(8):1057-1065. doi: 10.1111/1756-185X.13898. Epub 2020 Jun 29.
Osteoarthritis (OA) is a prevalent chronic diseases characterized by formation of osteophytes and degradation of articular cartilage. Previous evidence has identified the regulatory effects of microRNAs (miRNAs) in OA. The goal of this study is to clearly explore the biological function of miR-1207-5p in OA.
MiR-1207-5p and C-X3-C motif chemokine receptor 1 (CX3CR1) expression in OA cartilages were revealed by accessing to Gene Expression Omnibus database. In vitro OA model was established by lipopolysaccharide (LPS) stimulation. Western blot and quantitative real-time polymerase chain reaction were conducted to detect the expression level of genes. Cell counting kit-8 (CCK-8) and flow cytometric experiments were performed to investigate the proliferation and apoptosis capacities of CHON-001 cells. Bioinformatics analysis was applied to predict the binding site of miR-1207-5p and CX3CR1, the connections of which were ascertained using luciferase reporter assay.
MiR-1207-5p expression was decreased while CX3CR1 was increased in OA cartilages. Up-regulation of miR-1207-5p alleviated the LPS-induced damage in the view of cell proliferation, apoptosis and extracellular matrix (ECM) degradation. A target of miR-1207-5p CX3CR1, its down-regulation intensified the impacts of miR-1207-5p mimic, promoted proliferation and mitigated apoptosis. LPS exposure increased the protein expression of the phosphorylated IκBα and P65, and this phenomena was reversed due to miR-1207-5p up-regulation and CX3CR1 knockdown. The treatment of Betulinic acid (BA; an activator of nuclear factor-κB pathway) reversed the miR-1207-5p mimic-induced inhibitory effect on apoptosis in LPS-treated CHON-001.
Our results highlight that miR-1207-5p can prevent CHON-001 from LPS-stimulated injury, providing a novel biomarker for OA progression and further advancing treatment of OA.
骨关节炎(OA)是一种常见的慢性疾病,其特征为骨赘形成和关节软骨降解。先前的证据已经确定了 microRNAs(miRNAs)在 OA 中的调节作用。本研究的目的是明确探讨 miR-1207-5p 在 OA 中的生物学功能。
通过访问基因表达综合数据库揭示 OA 软骨中的 miR-1207-5p 和 C-X3-C 基序趋化因子受体 1(CX3CR1)表达。通过脂多糖(LPS)刺激建立体外 OA 模型。通过 Western blot 和定量实时聚合酶链反应检测基因的表达水平。通过细胞计数试剂盒-8(CCK-8)和流式细胞术实验研究 CHON-001 细胞的增殖和凋亡能力。应用生物信息学分析预测 miR-1207-5p 和 CX3CR1 的结合位点,通过荧光素酶报告基因实验确定它们的联系。
OA 软骨中 miR-1207-5p 表达降低,而 CX3CR1 表达增加。miR-1207-5p 的上调减轻了 LPS 诱导的细胞增殖、凋亡和细胞外基质(ECM)降解损伤。miR-1207-5p 的靶标 CX3CR1,其下调增强了 miR-1207-5p 模拟物的影响,促进了增殖并减轻了凋亡。LPS 暴露增加了磷酸化 IκBα 和 P65 的蛋白表达,而由于 miR-1207-5p 的上调和 CX3CR1 的敲低,这种现象被逆转。BA(核因子-κB 途径的激活剂)的处理逆转了 miR-1207-5p 模拟物在 LPS 处理的 CHON-001 中对凋亡的抑制作用。
我们的结果强调了 miR-1207-5p 可以防止 CHON-001 受到 LPS 刺激的损伤,为 OA 进展提供了一种新的生物标志物,并进一步推进了 OA 的治疗。